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Updated: Jan 31, 2026
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The Production of C. elegans Transgenes via Recombineering with the galK Selectable Marker
Published on: January 11, 2011
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New Selectable Markers for Volvox carteri Transformation.
José A Ortega-Escalante1, Owen Kwok1, Stephen M Miller1
1Department of Biological Sciences, University of Maryland Baltimore County, 1000 Hilltop Circle, Baltimore, MD 21250, USA.
Protist
|December 22, 2018
Summary
New selectable markers for Volvox carteri, a model organism for multicellularity, were developed. These markers provide antibiotic resistance, enabling efficient genetic manipulation and co-transformation for future research.
Area of Science:
- Evolutionary Biology
- Developmental Biology
- Molecular Genetics
Background:
- Volvox carteri is a key model organism for studying the evolution of multicellularity and cell differentiation.
- Existing molecular genetic tools for V. carteri are limited, hindering further progress.
- There is a need for additional selectable markers to enable multiplexing of transgenes.
Purpose of the Study:
- To develop novel selectable markers for efficient nuclear transformation in Volvox carteri.
- To facilitate advanced molecular genetic analyses and gene manipulation in V. carteri.
Main Methods:
- Development of constructs containing Volvox-specific regulatory sequences and codon-optimized antibiotic resistance genes (hygromycin and blasticidin).
- Transformation of V. carteri using biolistic bombardment with the new constructs.
- Selection of antibiotic-resistant transformants and detection of transgenes and transcripts.
Main Results:
- Two new selectable markers, VcHyg (hygromycin resistance) and VcBlast (blasticidin resistance), were successfully generated.
- Transformants were obtained at rates of 0.5-13 per million cells, with survivors readily isolated within 7 days.
- Co-transformation rates using the new markers were comparable to the established nitA marker.
Conclusions:
- The developed pVcHyg and pVcBlast plasmids are efficient and convenient tools for transforming Volvox carteri.
- These markers facilitate the multiplexing of transgenes, significantly advancing molecular genetic studies in V. carteri.
- The new markers support a broad range of V. carteri strains, enhancing their utility in research.
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